大型反应器内pH不均一性对CHO细胞流加培养过程的影响

doi:10.13560/j.cnki.biotech.bull.1985.2015.10.003

生物技术通报 ›› 2015, Vol. 31 ›› Issue (10): 236-241.doi: 10.13560/j.cnki.biotech.bull.1985.2015.10.003

大型反应器内pH不均一性对CHO细胞流加培养过程的影响

刘金涛¹,王星懿¹,范里¹,邓献存^1,2,刘旭平¹,谭文松¹

1.华东理工大学生物反应器国家重点实验室, 上海 200237;
2.浙江海正药业股份有限公司, 台州 318000

收稿日期:2015-01-09 出版日期:2015-10-28 发布日期:2015-10-28
作者简介:刘金涛, 男, 博士, 研究方向：动物细胞与组织工程; E-mail：jtliu0416@gmail.com
基金资助:
国家自然科学基金项目（21106045）

Effect of pH Heterogeneity in Large-scale Bioreactor on Fed-batch Culture Process of CHO cells

Liu Jintao¹, Wang Xingyi¹, Fan Li¹, Deng Xiancun^{1, 2}, Liu Xuping¹, Tan Wensong¹

1.State Key Lab of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237; 2. Hisun Pharmaceutical Co.Ltd, Taizhou 318000

Received:2015-01-09 Published:2015-10-28 Online:2015-10-28

摘要/Abstract

摘要： 为了研究大型反应器中pH不均一性对CHO细胞流加培养过程的影响, 并将培养过程顺利地放大到生产规模, 根据大型反应器的混合特性, 构建了搅拌式反应器与平推流反应器串联的规模缩小装置用于模拟大型反应器中的pH不均一性。结果表明停留时间为30 s时, 整个培养过程和对照相比并无显著的差异, 这表明此时补碱所导致的pH不均一性并未对流加培养工艺造成影响。而随着停留时间的延长, 反应器内pH不均一的程度越大, 细胞生长和产物表达受到抑制越明显; 与此同时, 乳酸和氨的累积显著增加, 而关键质量属性唾液酸和生物学活性也随之降低。

关键词: CHO细胞, 规模缩小, pH不均一性, TNFR-Fc, 反应器放大

Abstract: In order to study the effect of pH heterogeneity in large-scale bioreactor on cell culture process of CHO cells, we established a scale down model consistent of stirred tank reactor and plug flow reactor to simulate the pH heterogeneity of large-scale bioreactor based on the mixing characteristic. The results showed that the scale down process with 30 s residence time has no statically difference with the control process. However, significant effect on cell growth, cell metabolism and protein production were found when increased the residence time of PFR. Cell growth rate decreased accompanied by tremendously increase of ammonia and lactate when increased the pH heterogeneity. In addition, the titer, sialic acid content and bioactivity of antibody fusion protein were also decreased when increased the pH heterogeneity.

Key words: CHO cells, scale down, pH heterogeneity, TNFR-Fc, scale up

刘金涛,王星懿,范里,邓献存,刘旭平,谭文松. 大型反应器内pH不均一性对CHO细胞流加培养过程的影响[J]. 生物技术通报, 2015, 31(10): 236-241.

Liu Jintao, Wang Xingyi, Fan Li, Deng Xiancun, Liu Xuping, Tan Wensong. Effect of pH Heterogeneity in Large-scale Bioreactor on Fed-batch Culture Process of CHO cells[J]. Biotechnology Bulletin, 2015, 31(10): 236-241.

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大型反应器内pH不均一性对CHO细胞流加培养过程的影响

Effect of pH Heterogeneity in Large-scale Bioreactor on Fed-batch Culture Process of CHO cells

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